Jhample Sowmya B, Bhagwat Prashant K, Nerlekar Nisha A, Nandre Vinod S, Dandge Padma B
Department of Biochemistry, Shivaji University, Kolhapur, 416004, M.S., India.
Department of Microbiology, Shivaji University, Kolhapur, 416004, M.S., India.
Curr Microbiol. 2025 Jul 26;82(9):412. doi: 10.1007/s00284-025-04398-5.
The most common cause of cardiovascular diseases which impact blood arteries is thrombosis. Current medications and protein-based treatments are examples of thrombolytic agents. Finding new enzymatic sources is essential in health difficulties because all of these agents have unfavorable side effects. By taking this fact into account, this study presents the purification and biochemical characterization of a fibrinolytic metalloprotease from Proteus penneri SP-20 (AB909494) as a new isolate. The enzyme was successfully purified using DEAE-cellulose anion exchange chromatography at a recovery rate of 58.73% and at 33.38 purification fold. The SDS-PAGE examination yielded a molecular mass of 16 kDa. The isolated enzyme was most active at 40 °C and pH 6.0. The purified enzyme maximal velocity (Vmax) and Michaelis constant (Km) were determined to be 6 × 10-4 mol/L/min and 2.07 mg/mL, respectively. Phenanthroline, EDTA, Hg⁺, Ag⁺, Pb⁺, Li⁺, and Co⁺ all significantly lowered enzyme activity (≥ 70%), but Mn⁺, Fe⁺, Triton X-100, and urea markedly increased it. The enzyme showed considerable stability in a variety of laundry detergents and stability in toluene and hexane. The purified enzyme was highlighted as a potential safe thrombolytic drug by hemolytic experiments, which verified its non-hemolytic nature. These results imply that the enzymes have lots of potential for use in a range of industrial and medicinal applications.
影响血管的心血管疾病最常见的病因是血栓形成。当前的药物和基于蛋白质的治疗方法都是溶栓剂的例子。由于所有这些药物都有不良副作用，因此寻找新的酶源对于解决健康难题至关重要。考虑到这一事实，本研究展示了从奇异变形杆菌SP-20（AB909494）中分离出的一种新型纤溶金属蛋白酶的纯化及生化特性。该酶通过DEAE-纤维素阴离子交换色谱法成功纯化，回收率为58.73%，纯化倍数为33.38。SDS-PAGE检测得出其分子量为16 kDa。分离出的酶在40°C和pH 6.0时活性最高。纯化酶的最大反应速度（Vmax）和米氏常数（Km）分别测定为6×10-4 mol/L/min和2.07 mg/mL。邻菲罗啉、EDTA、Hg⁺、Ag⁺、Pb⁺、Li⁺和Co⁺均显著降低酶活性（≥70%），但Mn⁺、Fe⁺、Triton X-100和尿素则显著提高酶活性。该酶在多种洗衣粉中表现出相当的稳定性，在甲苯和己烷中也具有稳定性。溶血实验表明纯化酶具有非溶血特性，从而突出了其作为潜在安全溶栓药物的特性。这些结果表明，该酶在一系列工业和医学应用中具有很大的应用潜力。
